Lactoferrin and brain health and development in infants

ABSTRACT

The present invention relates generally to the field of brain development and brain health. One embodiment of the present invention relates to a composition that can be used for the treatment or prevention of a delayed brain development and/or a delayed development of the nervous system. Also cognitive performance can be increased.

CROSS REFERENCE TO RELATED APPLICATIONS

The present application is a National Stage of International ApplicationNo. PCT/EP2010/056237, filed on May 7, 2010, which claims priority toEuropean Patent Application No. 09159966.2, filed on May 12, 2009, theentire contents of which are being incorporated herein by reference.

The present invention relates generally to the field of brain health,brain protection and brain development. One embodiment of the presentinvention relates to a composition that can be used for the treatment orprevention of a delayed brain development and/or a delayed developmentof the nervous system. Neuronal cells in the brain can be protected.Also cognitive performance can be increased.

Mother's milk is recommended for all infants. However, in some casesbreast feeding is inadequate or unsuccessful or inadvisable for medicalreasons or the mother chooses not to breast feed either at all or for aperiod of more than a few weeks. Infant feeding formulas have beendeveloped for these situations. Infant feeding formulas are commonlyused today to provide supplemental or sole source nutrition early inlife. They may be used instead of or in addition to mother's milk tofeed infants. Consequently, they are often designed today to resemblemother's milk as closely as possible in terms of composition andfunction.

Recently, evidence is accumulating that breastfeeding may providelong-term cognitive advantages, particularly for infants born small orpremature (Anderson et al., Am J Clin Nutr 1999; 70:525-35; Lucas etal., Lancet 1992; 339:261-4). However, the underlying mechanism toexplain the relationship between breastfeeding and cognitive developmentremains unclear.

It has been suggested that docohexanoic acid (DHA) and arachidonic acid(AA) that are present in human milk might play a role in the observedeffect.

A further study claims that dietary sialic acid supplementation improveslearning and memory in piglets (Wang et al., 2007, American Journal ofClinical Nutrition, Vol. 85, No. 2, 561-569). Sialic acid is known to bea key component of both human milk oligosaccharides and neural tissues.

The nervous system is a highly complex network composed of neuronal andglial cells. It is present in all mammalian species. The nervous systemis composed of the central nervous system (brain and spinal cord) andthe peripheral nervous system (somatic, autonomous and enteric nervoussystem). The central nervous system drives the cognitive functions(memory, attention, perception, action, etc). Together with theperipheral nervous system, it has a fundamental role in the control ofbehaviour.

The nervous system develops during gestation and then refines to amature, functional network during the post natal period. Immaturity ordelayed maturation of the nervous system leads to delayed establishmentof the important biological functions that it regulates. For example,this may lead to a delayed establishment of cognitive functions(learning ability, attention, etc).

It is commonly accepted that cognitive development and cognitiveperformance have a significant influence on the quality of life. Itwould hence be desirable to have available a composition that allowssupporting the development and performance of the nervous system and ofthe brain.

It was consequently an object of the present invention to improve thestate of the art and to provide a composition that is based on naturalingredients and that allows supporting the development and performanceof the nervous system and of the brain, in particular of infants, forexample IUGR infants.

This object was achieved by the subject matter of the independentclaims.

The present inventors were able to demonstrate that lactoferrin, forexample a composition supplemented with lactoferrin, can be used toimprove brain development and cognitive functioning.

It could further be shown that the administration of lactoferrin allowsit to increase the neuron density and neuron survival.

Metabolic changes in the hippocampus measured by MRS after lactoferrinadministration imply that learning and short term memory is modulated.

Metabolic changes in the cortex after lactoferrin administration implythat long term memory is modulated.

Lactoferrin (LF), also known as lactotransferrin (LTF), is a globularmultifunctional protein that is known to exhibit an antimicrobialactivity and is a part of the innate defense, mainly at mucoses.

Lactoferrin may be found for example in milk and whey and in manymucosal secretions such as tears and saliva. As such, Lactoferrin may bepurified, e.g., from milk or may be produced recombinantly.

The present invention relates to lactoferrin obtainable from any source.

Lactoferrin from milk or whey, for example, has the advantage that it isa natural ingredient obtained from a food-grade composition and canconsequently be used as enriched fraction of the food compositionwithout further purification.

Recombinantly obtained lactoferrin has the advantage that it can beproduced easily in high concentrations.

Human colostrum has a relatively high concentration of lactoferrin,followed by human milk, then cow milk.

The composition of the present invention can be in particular effectivein IUGR mammals. Intrauterine Growth Restriction (IUGR) is a term usedto describe a condition in which the fetus or infant is smaller thanexpected for the number of weeks of pregnancy. A fetus or infant withIUGR often has a weight that is reduced by at least 10% compared tonormal fetusses or infants of the same gestational age. For example ahuman fetus with IUGR may be born at term (after 37 weeks of pregnancy)or prematurely (before 37 weeks).

The present inventors have found that lactoferrin or lactoferrinenriched compositions may be used to protect neuronal cells againstdegeneration. Such a degeneration may follow, for example, stress.Lactoferrin was found to promote neuronal survival and/or limit orprevent neuronal death in the brain.

In infants the lactoferrin and/or the lactoferrin containingcompositions of the present inventions may be used to protect thecentral nervous system from any stress occurring during the neuronaldevelopment period, and—consequently—to limit and/or preventstress-induced neuronal growth retardation and associated cognitivedysfunctions.

For the purpose of the present invention, the term “infant” includeschildren and comprises subjects in the age range from 0-14 years.

A human infant less than a month old is a newborn or a neonate. The term“newborn” includes premature infants, postmature infants and full termnewborns. Upon reaching the age of one or beginning to walk, infants arealso referred to as “toddlers” (generally 12-36 months).

Lactoferrin and/or the composition of the present invention may beadministered for example to

-   -   premature or term-born infants having experienced an        intrauterine growth retardation that may occur following any        adverse events during the gestation (smoking of the mother,        medication of the mother, low placenta quality, abnormal        placenta positioning, malnutrition of the mother and the fetus,        etc)    -   Premature infants without any intrauterine growth retardation    -   Very low/low birth weight infants    -   IUGR infants    -   Neonates and children showing brain growth retardation following        for example hypoxemia-ischemia at birth or any other adverse        event    -   Neonates and children showing cognitive disfunctions,        retardation

Lactoferrin or the composition of the present invention can therefore beadministered to the infant and/or to the mother during the gestationand/or lactation period.

Consequently, one embodiment of the present invention is an ingestiblecomposition enriched in lactoferrin.

Enriched means that lactoferrin was either added to the composition, sothat the resulting lactoferrin content of the composition is higher thanthe lactoferrin content of the composition without lactoferrin addition,or that the composition was treated in a way to concentrate the naturallactoferrin content in a composition.

Lactoferrin may also be provided as pure compound.

Alternatively, lactoferrin may be provided as a lactoferrin enrichedfraction, for example a lactoferrin enriched milk or whey fraction.

As milk or whey source bovine milk, human milk, goat milk, camel milk,horse milk and/or donkey milk may be used, for example. Colostrum may beused as well.

In therapeutic applications, compositions are administered in an amountsufficient to at least partially cure or arrest the symptoms of thedisorder and/or its complications. An amount adequate to accomplish thisis defined as “a therapeutically effective dose”. Amounts effective forthis purpose will depend on a number of factors known to those of skillin the art such as the severity of the disorder and the weight andgeneral state of the patient. In prophylactic applications, compositionsaccording to the invention are administered to a patient susceptible toor otherwise at risk of a particular disorder in an amount that issufficient to at least partially reduce the risk of developing adisorder. Such an amount is defined to be “a prophylactic effectivedose”. Again, the precise amounts depend on a number of patient specificfactors such as the patient's state of health and weight.

Lactoferrin may be administered in the framework of the presentinvention in a therapeutically effective dose and/or in a prophylacticeffective dose.

Typical lactoferrin enriched compositions may comprise lactoferrin in anamount of at least 1.6 g/L.

For example, the composition of the present invention may containlactoferrin in a concentration of at least 0.75% (w/w), preferably atleast 1% (w/w).

In one embodiment, the composition is to be administered in an amountcorresponding to an ingestion of at least 0.25 g lactoferrin, preferablyat least 0.5 g lactoferrin more preferably at least 1 g lactoferrin perday per kg body weight.

For example, the composition may be consumed in an amount correspondingto at least 1 g lactoferrin/kg body weight/day intake for pregnantand/or lactating mothers.

The composition may also be consumed in an amount corresponding to atleast 200 mg lactoferrin/kg body weight/day intake for the children.

Lactoferrin may be present in the composition in a concentration of atleast 0.01 g per 100 kcal, preferably of at least 0.1 g per 100 kcal.For example, lactoferrin may be present in the composition in the rangeof about 0.01 g-100 g, preferably 0.1 g-50 g, even more preferred 2 g-25g per 100 kcal of the composition.

Lactoferrin may also be used in combination with other compounds, suchas sialic acid and/or iron, for example.

A particular preferred lactoferrin containing composition may containadditionally sialic acid in an amount in the range of 100 mg/100 g (w/w)to 1000 mg/100 g (w/w) of the composition, for example in the range of500 mg/100 g (w/w) to 650 mg/100 g (w/w) of the composition.

The composition of the present invention may for example comprise atleast about 0.001 weight-% sialic acid. In further embodiments of thepresent invention, the composition may comprise at least about 0.005weight-%, or at least about 0.01 weight-% of sialic acid.

Alternatively or additionally the lactoferrin containing composition maycontain iron in an amount in the range of about 1 mg/100 g (w/w) to 50mg/100 g (w/w) of the composition, for example 10 mg/100 g (w/w) to 30mg/100 g (w/w) of the composition.

One lactoferrin containing composition may contain for example about 852mg/100 g (w/w) sialic acid and 22 mg/100 g (w/w) iron.

The lactoferrin containing composition of the present invention may havea caloric density in the range of 30 kcal/100 g-1000 kcal/100 g of thecomposition, preferably 50 kcal/100 g-450 kcal/100 g of the composition.It may for example have a caloric density of about 400 kcal/100 g.

The nature of the composition is not particularly limited. It ispreferably a composition for oral or enteral administration.

The composition may be for example selected from the group consisting offood products, animal food products, pharmaceutical compositions,nutritional formulations, nutraceuticals, drinks, food additives, andinfant feeding formulas.

In one typical embodiment of the present invention, the composition willcontain a protein source, a lipid source and a carbohydrate source.

For example such a composition may comprise protein in the range ofabout 2 to 6 g/100 kcal, lipids in the range of about 1.5 to 3 g/100kcal and/or carbohydrates in the range of about 1.7 to 12 g/100 kcal

If the composition is liquid, its energy density may be between 60 and75 kcal/100 ml.

If the composition is solid, its energy density may be between 60 and 75kcal/100 g.

The type of protein is not believed to be critical to the presentinvention. Thus, protein sources based on whey, casein and mixturesthereof may be used, for example. As far as whey proteins are concerned,acid whey or sweet whey or mixtures thereof may be used as well asalpha-lactalbumin and beta-lactoglobulin in whatever proportions aredesired. The whey protein may be modified sweet whey. Sweet whey is areadily available by-product of cheese making and is frequently used inthe manufacture of infant formulas based on cows' milk. However, sweetwhey includes a component which is undesirably rich in threonine andpoor in tryptophan called caseino-glyco-macropeptide (CGMP). Removal ofthe CGMP from sweet whey results in a protein with a threonine contentcloser to that of human milk. This modified sweet whey may then besupplemented with those amino acids in respect of which it has a lowcontent (principally histidine and tryptophan). A process for removingCGMP from sweet whey is described in EP 880902 and an infant formulabased on this modified sweet whey is described in WO 01/11990. Theproteins may be intact or hydrolysed or a mixture of intact andhydrolysed proteins. It may be desirable to supply partially hydrolysedproteins (degree of hydrolysis between 2 and 20%), for example forsubjects believed to be at risk of developing cows' milk allergy. Ifhydrolysed proteins are required, the hydrolysis process may be carriedout as desired and as is known in the art. For example, a whey proteinhydrolysate may be prepared by enzymatically hydrolysing the wheyfraction in two steps as described in EP 322589. For an extensivelyhydrolysed protein, the whey proteins may be subjected to triplehydrolysis using Alcalase 2.4 L (EC 940459), then Neutrase 0.5 L(obtainable from Novo Nordisk Ferment AG) and then pancreatin at 55° C.If the whey fraction used as the starting material is substantiallylactose free, it is found that the protein suffers much less lysineblockage during the hydrolysis process. This enables the extent oflysine blockage to be reduced from about 15% by weight of total lysineto less than about 10% by weight of lysine; for example about 7% byweight of lysine which greatly improves the nutritional quality of theprotein source.

The compositions of the present invention may contain a carbohydratesource. Any carbohydrate source may be used, such as lactose,saccharose, maltodextrin, starch and mixtures thereof.

The compositions of the present invention may contain a lipid source.The lipid source may be any lipid. Preferred fat sources include milkfat, palm olein, high oleic sunflower oil and high oleic safflower oil.The essential fatty acids linoleic and α-linolenic acid may also beadded as may small amounts of oils containing high quantities ofpreformed arachidonic acid and docosahexaenoic acid such as fish oils ormicrobial oils. The lipid source preferably has a ratio of n-6 to n-3fatty acids of about 5:1 to about 15:1; for example about 8:1 to about10:1.

The compositions of the present invention may also contain all vitaminsand minerals understood to be essential in the daily diet and innutritionally significant amounts.

Minimum requirements have been established for certain vitamins andminerals. Examples of minerals, vitamins and other nutrients optionallypresent in the infant formula include vitamin A, vitamin B1, vitamin B2,vitamin B6, vitamin B12, vitamin E, vitamin K, vitamin C, vitamin D,folic acid, inositol, niacin, biotin, pantothenic acid, choline,calcium, phosphorous, iodine, iron, magnesium, copper, zinc, manganese,chloride, potassium, sodium, selenium, chromium, molybdenum, taurine,and L-carnitine. Minerals are usually added in salt form. The presenceand amounts of specific minerals and other vitamins will vary dependingon the numerous factors, such as age weight and condition of the personor animal the composition is administered to.

The compositions may also comprise at least one probiotic bacterialstrain. A probiotic is a microbial cell preparation or components ofmicrobial cells with a beneficial effect on the health or well-being ofthe host. Suitable probiotic bacterial strains include Lactobacillusrhamnosus ATCC 53103 obtainable from Valio Oy of Finland under the trademark LGG, Lactobacillus rhamnosus CGMCC 1.3724, Lactobacillus paracaseiCNCM I-2116, Lactobacillus reuteri ATCC 55730 and Lactobacillus reuteriDSM 17938 obtainable from BioGaia AB, Bifidobacterium lactis CNCM I-3446sold inter alia by the Christian Hansen company of Denmark under thetrade mark Bb12 and Bifidobacterium longum ATCC BAA-999 sold by MorinagaMilk Industry Co. Ltd. of Japan under the trade mark BB536. The amountof probiotic, if present, likewise preferably varies as a function ofthe age of the person or animal. Generally speaking, the probioticcontent may increase with increasing age of the infant for example from10³ to 10¹² cfu/g formula, more preferably between 10⁴ and 10⁸ cfu/gformula (dry weight).

The compositions may also contain at least one prebiotic in an amount of0.3 to 10%. A prebiotic is a non-digestible food ingredient thatbeneficially affects the host by selectively stimulating the growthand/or activity of one or a limited number of bacteria in the colon, andthus improves host health. Such ingredients are non-digestible in thesense that they are not broken down and absorbed in the stomach or smallintestine and thus pass intact to the colon where they are selectivelyfermented by the beneficial bacteria. Examples of prebiotics includecertain oligosaccharides, such as fructooligosaccharides (FOS) andgalactooligosaccharides (GOS). A combination of prebiotics may be usedsuch as 90% GOS with 10% short chain fructo-oligosaccharides such as theproduct sold under the trade mark Raftilose® or 10% inulin such as theproduct sold under the trade mark Raftiline®.

A particularly preferred prebiotic is a mixture ofgalacto-oligosaccharide(s), N-acetylated oligosaccharide(s) andsialylated oligosaccharide(s) in which the N-acetylatedoligosaccharide(s) comprise 0.5 to 4.0% of the oligosaccharide mixture,the galacto-oligosaccharide(s) comprise 92.0 to 98.5% of theoligosaccharide mixture and the sialylated oligosaccharide(s) comprise1.0 to 4.0% of the oligosaccharide mixture. This mixture is hereinafterreferred to as “CMOS-GOS”. Preferably, a composition for use accordingto the invention contains from 2.5 to 15.0 wt % CMOS-GOS on a dry matterbasis with the proviso that the composition comprises at least 0.02 wt %of an N-acetylated oligosaccharide, at least 2.0 wt % of agalacto-oligosaccharide and at least 0.04 wt % of a sialylatedoligosaccharide.

Suitable N-acetylated oligosaccharides include GalNAcα1,3Galβ1,4Glc andGalβ1,6GalNAcα1,3Galβ1,4Glc. The N-acetylated oligosaccharides may beprepared by the action of glucosaminidase and/or galactosaminidase onN-acetyl-glucose and/or N-acetyl galactose. Equally, N-acetyl-galactosyltransferases and/or N-acetyl-glycosyl transferases may be used for thispurpose. The N-acetylated oligosaccharides may also be produced byfermentation technology using respective enzymes (recombinant ornatural) and/or microbial fermentation. In the latter case the microbesmay either express their natural enzymes and substrates or may beengineered to produce respective substrates and enzymes. Singlemicrobial cultures or mixed cultures may be used. N-acetylatedoligosaccharide formation can be initiated by acceptor substratesstarting from any degree of polymerisation (DP) from DP=1 onwards.Another option is the chemical conversion of keto-hexoses (e.g.fructose) either free or bound to an oligosaccharide (e.g. lactulose)into N-acetylhexosamine or an N-acetylhexosamine containingoligosaccharide as described in Wrodnigg, T. M.; Stutz, A. E. (1999)Angew. Chem. Int. Ed. 38:827-828. Suitable galacto-oligosaccharidesinclude Galβ1,6Gal, Galβ1,6Galβ1,4Glc Galβ1,6Galβ1,6Glc,Galβ1,3Galβ1,3Glc, Galβ1,3Galβ1,4Glc, Galβ1,6Galβ1,6Galβ1,4Glc,Galβ1,6Galβ1,3Galβ1,4Glc Galβ1,3Galβ1,6Galβ1,4Glc,Galβ1,3Galβ1,3Galβ1,4Glc, Galβ1,4Galβ1,4Glc andGalβ1,4Galβ1,4Galβ1,4Glc.

Synthesised galacto-oligosaccharides such as Galβ1,6Galβ1,4GlcGalβ1,6Galβ1,6Glc, Galβ1,3Galβ1,4Glc, Galβ1,6Galβ1,6Galβ1,4Glc,Galβ1,6Galβ1,3Galβ1,4Glc and Galβ1,3Galβ1,6Galβ1,4Glc, Galβ1,4Galβ1,4Glcand Galβ1,4Galβ1,4Galβ1,4Glc and mixtures thereof are commerciallyavailable under the trade marks Vivinal® and Elix'or®. Other suppliersof oligosaccharides are Dextra Laboratories, Sigma-Aldrich Chemie GmbHand Kyowa Hakko Kogyo Co., Ltd. Alternatively, specificglycoslytransferases, such as galactosyltransferases may be used toproduce neutral oligosaccharides.

Suitable sialylated oligosaccharides include NeuAcα2,3Galβ1,4Glc andNeuAcα2,6Galβ1,4Glc. These sialylated oligosaccharides may be isolatedby chromatographic or filtration technology from a natural source suchas animal milks. Alternatively, they may also be produced bybiotechnology using specific sialyltransferases either by enzyme basedfermentation technology (recombinant or natural enzymes) or by microbialfermentation technology. In the latter case microbes may either expresstheir natural enzymes and substrates or may be engineered to producerespective substrates and enzymes. Single microbial cultures or mixedcultures may be used. Sialyl-oligosaccharide formation can be initiatedby acceptor substrates starting from any degree of polymerisation (DP)from DP=1 onwards.

The compositions may optionally contain other substances which may havea beneficial effect such as nucleotides, nucleosides, and the like.

The compositions, for example an infant formula, for use in theinvention may be prepared in any suitable manner. For example, an infantformula may be prepared by blending together the protein source, thecarbohydrate source, and the fat source in appropriate proportions. Ifused, the emulsifiers may be included in the blend. The vitamins andminerals may be added at this point but are usually added later to avoidthermal degradation. Any lipophilic vitamins, emulsifiers and the likemay be dissolved into the fat source prior to blending. Water,preferably water which has been subjected to reverse osmosis, may thenbe mixed in to form a liquid mixture. The liquid mixture may then bethermally treated to reduce bacterial loads. For example, the liquidmixture may be rapidly heated to a temperature in the range of about 80°C. to about 110° C. for about 5 seconds to about 5 minutes. This may becarried out by steam injection or by heat exchanger; for example a plateheat exchanger. The liquid mixture may then be cooled to about 60° C. toabout 85° C.; for example by flash cooling. The liquid mixture may thenbe homogenised; for example in two stages at about 7 MPa to about 40 MPain the first stage and about 2 MPa to about 14 MPa in the second stage.The homogenised mixture may then be further cooled to add any heatsensitive components; such as vitamins and minerals. The pH and solidscontent of the homogenised mixture is conveniently standardised at thispoint. The homogenised mixture is transferred to a suitable dryingapparatus such as a spray drier or freeze drier and converted to powder.The powder should have a moisture content of less than about 5% byweight. If it is desired to add probiotic(s), they may be culturedaccording to any suitable method and prepared for addition to the infantformula by freeze-drying or spray-drying for example. Alternatively,bacterial preparations can be bought from specialist suppliers such asChristian Hansen and Morinaga already prepared in a suitable form foraddition to food products such as infant formula. Such bacterialpreparations may be added to the powdered infant formula by dry mixing.

Lactoferrin may be added at any stage during this procedure, but ispreferably added after the heating step.

The composition comprises a protein source which may be present in therange of between 1.4 and 100 g/100 kcal, preferably between 1.4 and 6.0g/100 kcal of the composition. Since lactoferrin is a protein it shouldbe considered a part of the protein source.

Whey protein is known to provide several health benefits. For example,it is easily digestible. The protein fraction in whey (approximately 10%of the total dry solids within whey) comprises several proteinfractions, for example beta-lactoglobulin, alpha-lactalbumin, bovineserum albumin and immunoglobulins. In one embodiment at least 50%,preferably at least 75%, even more preferred at least 85% by weight ofthe protein source is whey protein.

If present, the lipid source may contribute to between 30 to 55% of thetotal energy of the composition. A carbohydrate source may contribute tobetween 35 and 65% of the total energy of the composition.

Sialic acid may also be added to the composition of the presentinvention. Sialic acid is a generic term for the N- or O-substitutedderivatives of neuraminic acid, a monosaccharide with a nine-carbonbackbone.

Any sialic acid may be used for the purposes of the present invention.However, it is preferred if the sialic acid has the following formula

R1=H, acetyl, lactyl, methyl, sulfate, phosphate, anhydro, sialic acid,fucose, glucose, or galactose

R2=N-acetyl, N-glycolyl, amino, hydroxyl, N-glycolyl-O-acetyl, orN-glycolyl-O-methyl

R3=H, galactose, N-acetylglucosamine, N-acetylgalactosamine, sialicacid, or N-glycolylneuraminic acid

R1 may be selected from the group consisting of H, acetyl, lactyl,methyl, sulfate, phosphate, anhydrosialic acid, fucose, glucose and/orgalactose.

R2 may be selected from the group consisting of N-acetyl, N-glycolyl,amino, hydroxyl, N-glycolyl-O-acetyl, and/or N-glycolyl-O-methyl.

R3 may be selected from the group consisting of H, galactose,N-acetylglucosime, N-acetylgalactosamine, sialic acid, and/orn-glycolylneuraminic acid.

The groups in position R1 may be identical or may differ from eachother.

For example, the sialic acid may be N-acetylneuraminic acid with R1=H,R2=N-acetyl and R3=H. According to a further embodiment of the presentinvention the sialic acid may selected from the group consisting of2-keto-5-acetamido-3,5-dideoxy-d-glycero-d-galactononulosonic acid(Neu5Ac) and n-glycolylneuraminic acid or mixtures thereof.

Sialic acid as used in the present invention comprisesN-Acetylneuraminic acid, which has the following synonyms andabbreviations: o-Sialic acid;5-Acetamido-3,5-dideoxy-D-glycero-D-galacto-2-non ulosonic acid;5-Acetamido-3,5-dideoxy-D-glycero-D-galactonulosonic acid; Aceneuramicacid; N-acetyl-neuraminate; N-Acetylneuraminic acid; NANA, and Neu5Ac.

The present invention extends to the use of lactoferrin for thepreparation of a composition for the treatment or prevention of adelayed brain development and/or a delayed development of the nervoussystem.

For the uses of the present invention it is essential that thecomposition contains lactoferrin or a compound that yields lactoferrinafter consumption. The composition does not have to be enriched inlactoferrin, although this may be preferable, since this way morelactoferrin can be administered in smaller volumes.

The lactoferrin may be used to prepare any kind of composition. It ispreferred, however, that the lactoferrin is provided as a composition inaccordance with what is described above.

In one embodiment of the present invention, the lactoferrin containingcomposition may be used to treat or prevent a delayed visiondevelopment.

In another embodiment of the present invention, the lactoferrincontaining composition may be used to treat or prevent a delayed neuralmigration.

In a further embodiment of the present invention, the lactoferrincontaining composition may be used to treat or prevent a delayedcognitive development.

The composition of the present invention can be used to increase theneuronal density and or the neuronal survival.

The compositions of the present invention may further be used to treator prevent an impaired learning ability, an impaired mental performanceor a reduced attention span.

To achieve this, the composition may be administered to mothers duringpregnancy, mothers during lactation, to premature or term born babies,to very low/low birth weight infants, IUGR infants, infants, toddlers,children and/or teenagers.

While the compositions of the present invention can generally be used totreat or prevent brain disorders and/or to repair and/or reverse braindamage in infants of any age group, it was found that the compositionsof the present invention are particular helpful to treat or preventbrain disorders and/or to repair and/or reverse brain damage in IUGRinfants.

Those skilled in the art will understand that they can freely combineall features of the present invention described herein, withoutdeparting from the scope of the invention as disclosed. In particular,features described for the uses of the present invention may be appliedto composition of the present invention and vice versa.

Further advantages and features of the present invention are apparentfrom the following Examples and Figures.

FIG. 1 shows the percentage of positive NS20Y cells for neuriteoutgrowth in basal condition (untreated cells) and after treatment ofthe cells with either the neurotrophic factor CNTF (100 ng/mL, positivecontrol) or the lactoferrin enriched bovine milk fraction at differentconcentrations. Data are means±SEM, n=3 to 7 according to the group(Basal, n=7; CNTF, n=3; 1 ug/L, n=3; 10 ug/L, n=7; 100 ug/L, n=3; 1mg/L, n=3; 10 mg/L, n=5; 100 mg/L, n=7; 1 g/L, n=6). Data were comparedto the basal untreated group with the student t test. A difference wasconsidered significant when P<0.05.

FIG. 2 shows the release of neuron-specific enolase (NSE), a marker forneuronal cell death, by a primary culture of enteric neurons, followingH₂O₂ challenge and prevention with bovine milk Lactoferrin. Data aremean±SEM, n=8. A difference was considered significant when P<0.05

FIG. 3 shows the percentage of 7-AAD positive cells in cultured SH-SY5Ycells, following H₂O₂ challenge in presence or not of differentconcentrations of bovine milk Lactoferrin ranging from 0.001 to 1 g/L.

FIG. 4 a shows the brain to body weight ratio at P1 in normal and IUGRinfants. The brain to body weight ratio was found to be increased afterlactoferrin administration during gestation, both in normal infants andeven more in IUGR infants.

FIG. 4 b shows the brain to body weight ratio at P7 in normal and IUGRinfants. The brain to body weight ratio was found to be increased afterlactoferrin administration, both in normal infants and even more in IUGRinfants.

FIG. 5 shows the presence of several metabolic markers indicative forbrain activity and development in the hippocampus of normal infants,IUGR infants and IUGR infants treated with lactoferrin at P7.Hippocampal activity is linked to learning and short term memory.

FIG. 6 shows the presence of several metabolic markers indicative forbrain activity and development in the cortex of normal infants, IUGRinfants and IUGR infants treated with lactoferrin at P7. Cortex activityis linked to long term memory.

FIG. 7 shows the nuclei morphology in the CA2-CA3 field of thehippocampus after DAPI staining.

FIG. 8 shows that dietary lactoferrin supplementation significantlyincreased gene expression of brain derived neutrophic factor (BDNF) atpostnatal day 7.

EXAMPLES

Biological activity of lactoferrin enriched bovine milk fraction has aneffect on promoting neuronal cell survival and neurite outgrowth invitro

The neurite outgrowth process comprises the outgrowth of axons fromneurons and is part of neuronal development. The impact of a fraction ofbovine milk enriched in lactoferrin on neurite outgrowth was measuredusing a well established and commonly used in vitro bioassay.

Briefly, NS20Y murine neuroblastoma cells (DSMZ) were thawed fromcryogenic storage, plated at a density of approximately 27×10³ cells percm² in tissue culture-treated flasks (Falcon) and expanded in thepresence of DMEM (Gibco) containing 10% FCS (Gibco) and 2 mM L-glutamine(Gibco). Two days after plating, the cells were detached from the flaskby mechanical agitation (tapping of the flask), and a single cellsuspension was obtained by passing the suspension several times througha flame-polished glass pipette. Cells were then plated onto 13 mm roundglass coverslips in the presence of DMEM containing 10% FCS and 2 mML-glutamine at a density of 2,000 cells per coverslip. The following daythe medium was switched to DMEM containing 0.5% FCS, 2 mM L-glutamine,and different concentrations of the milk fractions to be tested. One daylater cells were fixed with 4% paraformaldehyde and the coverslipsmounted on slides.

All coverslips were imaged with an Axioplan 2 microscope (Zeiss).Digital images were taken from 25 defined fields across the diameter ofthe coverslip (20× objective, Axiocam MRc, Zeiss). Cells were countedsystematically from the first field at the edge of the coverslip acrossthe coverslip until 100 cells had been counted. Cells were scored foreither positive or negative for neurite outgrowth. Positive cells forneurite outgrowth were considered if the axon-resembling projectionsemanating from the cell body reached a length greater than the cellbody.

A student t test was used to compare differences in the mean between onecontrol reference population and means from all other treatments in eachgroup.

The following concentrations of the lactoferrin enriched bovine milkfraction were tested: 1 μg/L, 10 μg/L, 100 μg/L, 1 mg/L, 10 mg/L, 100mg/L, and 1 g/L. A positive control (CNTF, ciliary neurotrophic factor,100 ng/mL), which is a well known neurotrophic factor previouslyreported to promote neurite outgrowth of different neuronal populations(Oyesiku and Wigston, 1996 (Oyesiku N M, Wigston D J: Ciliaryneurotrophic factor stimulates neunte outgrowth from spinal cordneu-rons. J Comp Neurol 1996; 364: 68-77.) was performed. A basalcontrol consisted of untreated cells. Results are shown in FIG. 1.

Protection of Neuronal Cells Against Stress

Rat primary cultures of enteric neuronal cells were seeded into wellsand incubated with different concentrations of bovinelactoferrin-enriched fraction for 48 h. After washing three times withphosphate buffer saline (sterile PBS, 37° C.), the cells were incubatedfor 12 hours in cell medium without lactoferrin and containing H₂O₂ orits vehicle (control). The protective effect of lactoferrin uponH₂O₂-induced neuronal cell death was evaluated by measuring the releaseof neuron-specific enolase (NSE) in the cell medium. After oxidativestress, the medium of the different groups were collected andcentrifuged for 10 min at 12,000 rpm (4° C.). The supernatant wascollected and the NSE released in the culture medium was quantified byimmunoradiometric assay. Results are expressed in ng/mL. As shown onFIG. 2, H₂O₂ induced a significant increase of NSE in the medium(p<0.05, n=8). Treatment of primary neuronal enteric cells withlactoferrin-enriched fraction significantly prevented the H₂O₂-inducedrelease of NSE (p<0.05, n=8).

The neuroprotective property of bovine lactoferrin was confirmed using ahuman neuronal-like cell line (SH-SY5Y-neuroblastoma cells). Briefly,SH-SY5Y cells were plated for 24 h, and bovine lactoferrin-enrichedfraction was added to the culture media of cells at differentconcentrations for the following 48 h. Cells were challenged with H₂O₂for 6 h. Cells were finally washed with 0.1 M PBS before being harvestedwith trypsine-EDTA. Cell suspension was then pooled with the supernatantand centrifuged for 5 min at 2,000 rpm. After centrifugation, the pelletwas resuspended in 500 microliter of PBS 0.1 M. Membrane permeabilitywas evaluated by flow cytometry using the 7-AAD as fluorescent marker.For this, 200 microliter of cell suspension were incubated with 7-AADfor 10 min before acquisition using BD FACS Array™ bioanalyser. Thisflow cytometric assay using 7-aminoactinomycin D (7-AAD) allowed todistinguish live (7-AAD negative) and late apoptotic/necrotic (7-AADpositive) SH-SY5Y cells in response to oxidative stress. Results aspresented in FIG. 3 were expressed as percentage of 7-AAD positive cellsper total number of cells. As shown in FIG. 3, H₂O₂ induced asignificant increase in the percentage of 7-AAD positive cells (p<0.05,n=6). Treatment of SH-SY5Y cells with lactoferrin prevented theH₂O₂-induced increase in percentage of 7-ADD positive cells.

Lactoferrin Improves the Brain/Body Weight Ratio in Normal and IUGRInfants

Rat Model: Wister Rat

Dams are treated with dexamethasone (DEX) during the third week ofgestation. This corticosteroid will be delivered during the 3^(rd) ofgestation by an osmotic pump Alzet® embeded subcutaneously; sham animalswith osmotic pump containing saline buffer will be used as control. Thisdesign represents a model of high frailty for pups, mimicking asituation of frailty during the perinatal period in the human species,which is a property model to prove the ability of lactoferrin to enhancethe brain development apart from any other interventions. Lactoferrinsupplementation will be tested 1) during both gestation and lactation,2) during lactation and 3) no supplementation. For establishing alogical experimental design allowing proper comparisons, the samesupplementation protocol will be applied to normal gestations.

IUGR pups: model of intrauterine growth retardation (IUGR) is obtainedby the treatment of dams with dexamethasone (100 μg/kg/day) during thethird week of gestation. For the nutritional supplementation ofgestating dams, lactoferrin will be given orally from the 15^(th) day ofgestation to the weaning and food is available ad libitum. Lactoferrinwill be delivered to newborn rats from postnatal day 1 until they willbe weaned.

The following 6 groups of animals were used:

-   -   Group 1: Normal pups; no nutritional intervention in control        dams (sham=osmotic pump with saline buffer).    -   Group 2: IUGR pups; no nutritional intervention in DEX treated        dams.    -   Group 3: Normal pups; bLf supplementation (1 g/Kg/day) of        control dams (sham) from the beginning of gestation to the end        of the lactation.    -   Group 4: IUGR pups; bLf supplementation (1 g/Kg/day) of DEX        treated dams from the beginning of gestation to the end of the        lactation.    -   Group 5: IUGR pups; no nutritional intervention in DEX treated        dams; vehicule (same volume as bLf) of pups were drop fed 200        mg/kg/day of a blend of amino acids mimicking casein protein in        addition to lactation from 1 to 21 days after giving birth.    -   Group 6: IUGR pups; no nutritional intervention of DEX treated        dams; bLf supplementation (200 mg/Kg/day) of pups by drop        feeding in addition to lactation from 1 to 21 days after giving        birth.

The results were the following and are shown in FIG. 4 a) and b).

The body weight of the offspring at birth of the DEX control andlactoferrin supplementation DEX are about 20˜25% smaller than thatcontrol vehicule group. This shows that the DEX model is a valid tool tomimick a situation of frailty during the perinatal period in humanspecies.

Thus this model is a property model to demonstrate the ability oflactoferrin to enhance the brain development apart from any otherinterventions.

Brain weight in both DEX control and DEX lactoferrin supplementationgroups were smaller than that control vehicule group. However thedecrease of brain weight is smaller than that body weight in Lfsupplemented groups, thus brain to body weight ratio is bigger in Dex LFtreatment compared to the Dex control group at postnatal day 1.

Interestingly the brain weight of the Dex Lf group caught up to thecontrol vehicule group on postnatal day 21.

Lactoferrin Increases Metabolism in the Brain

Using LC model analysis, the following 18 metabolites will be quantifiedfrom both the cortex and hippocampus: alanine (Ala), aspartate (Asp),creatine (Cr), -aminobutyric acid (GABA), glucose (Glc), glutamate(Glu), glutamine (Gln), glutathione (GSH), glycerophosphorylcholine(GPC), phosphorylcholine (PCho), myo-inositol (Ins), lactate (Lac),N-acetylaspartate (NAA), N-acetylaspartylglutamate (NAAG),phosphocreatine (PCr), phosphorylethanolamine (PE), scyllo-inositol, andtaurine (Tau).

It was aimed to visualize changes in cerebral development followingadverse prenatal exposures using in vivo MR techniques (use of a 9.4Tesla scanner at the EPFL), and to assess the effect of earlynutritional interventions on brain development and metabolism mainlyduring the first month of life in our rodent models. Fast Spin-Echo(FSE) images and spectra edition ¹H-MR Spectroscopy were used for thespecific local cerebral and hippocampus metabolism. Briefly, FSE images(TR/TE=6000/80 ms; FOV=25×25 mm and matrix size=256×128) were realizedto position MRS voxel of interest (VOI=1.5×1.5×2.5 mm3). First andsecond order shims were adjusted using FASTMAP [Martin E, 2001, AnnNeurol 49:518-521]. The water linewidths ranged between 8 and 15 Hz.Spectra acquisitions both within the cortical lesion and thecontralateral cortical area were performed using an ultra-short echotime (TE/TR=2.7/4000 ms) SPECIAL spectroscopy method. This methodcombines 1D image-selected in vivo spectroscopy (ISIS) in the vertical(Y) direction with a slice selective spin echo in the X and Z directionsand provides full signal intensity available in the excited region. 35to 70 series of FIDs (12 averages each) were acquired, individuallycorrected for frequency drift, summed together and corrected forresidual eddy current effects using the reference water signal.

The results were the following and are shown in FIG. 5 and FIG. 6.

There are significant differences in phosphocreatine (PCr),N-acetylaspartylguatamate (NAAG), N-acetylaspartate (NAA), NAA+NAAG andcreatine (Cr)+phosphocreatine (PCr) concentration between controlvehicle (n=5) and control Dex pups (n=4) at 7 days after giving birth(P<0.05˜0.01). LF treatment Dex pup group (n=6) however, had a trend toreverse the concentrations of above metabolic markers found in Contr-Dexgroup at P7, but differences did not reach statistic significant in bothhippocampus and cortex.

N-Acetylaspartate (NAA), or N-acetylaspartic acid, is a derivative ofaspartic acid with a formula of C₆H₉NO₅ and a molecular weight of175.139. NAA is the second most concentrated molecule in the brain afterthe amino acid glutamate. NAA is synthesized in neurons from the aminoacid aspartate and acetyl coenzyme A. Its proposed primary functionsinclude:

-   -   it is a source of acetate for lipid and myelin synthesis in        oligodendrocytes, the glial cells that myelinate neuronal axons    -   it is a precursor for the synthesis of the important neuronal        dipeptide N-Acetylaspartylglutamate    -   it is a neuronal osmolyte that is involved in fluid balance in        the brain    -   NAA may also be involved in energy production from the amino        acid glutamate in neuronal mitochondria

The NAA signal reflects tissue concentrations of both NAA andN-acetylaspartylglutamate (NAAG). NAA has been reported to reflect thepresence of neurons, oligodendroglial lineage cells, and axons in theCNS (Urenjak J, 1993, J Neurosci 13:981-989; Martin E, 2001, Ann Neurol49:518-521; Bjartmar C, 2002, Ann Neurol 51:51-58). It has beensuggested that NAA(G) may be an acetyl-group carrier betweenmitochondria and cytoplasm in neuronal cells (Patel T B, 1979, Biochem J184:539-546; Truckenmiller M E, 1985, J Neurochem 45:1658-1662). Adecrease of the NAA signal is usually interpreted as a reduction in thenumber of neurons, but it may also reflect altered function of neuronalmitochondria. The increase of NAA/Cho ratios in cerebral tissue as aresult of maturation was previously described in detail and is confirmedin the present study (van der Knaap M S, 1990, Radiology 176:509-515;Kreis R, 2002, Magn Reson Med 48:949-958).

N-Acetylaspartylglutamic acid (N-acetylaspartylglutamate or NAAG) is aneuropeptide which is the third-most-prevalent neurotransmitter in themammalian nervous system. NAAG consists of N-acetylaspartic acid (NAA)and glutamic acid coupled via a peptide bond. NAAG was discovered as anervous system-specific peptide in 1965 by Curatolo and colleagues(Isaacks R E, 1994, Neurochem Res 19:331-338) but was not extensivelystudied for nearly 20 years. It meets the criteria for aneurotransmitter, including being concentrated in neurons, packed insynaptic vesicles, released in a calcium-dependent manner, andhydrolyzed in the synaptic space by enzymatic activity. NAAG activates aspecific receptor, the metabotropic glutamate receptor type 3. It issynthesized enzymatically from its two precursors and catabolized byNAAG peptidases in the synapse. The inhibition of the latter enzymes haspotentially important therapeutic effects in animal models of severalneurologic conditions and disorders.

myo-Inositol is a crucial constituent of living cells and participatesin several physiologic functions. It is a major osmolyte and also servesas the precursor to phosphatidylinositol. myo-inositol has been used asa glial cell marker (Isaacks R E, 1994, Neurochem Res 19:331-338). Lacmay be used as fuel for the brain but also for the synthesis of myelin(Sanchez-Abarca L I, 2001, Glia 36:321-329).

A decrease of the N-acetylaspartate/choline (NAA/Cho) ratio inasphyxiated full-term neonates predicts an adverse neurodevelopmentaloutcome (Groenendaal F, 1994, Pediatr Res 35:148-151; Peden C J, 1993,Dev Med Child Neurol 35:502-510; Roelants-van Rijn A M, 2001, PediatrRes 49:356-362). Myo-inositol (ml), which is one of the osmoregulatorsof the brain, can be found in astrocytes and is considered a glial cellmarker (Isaacks R E, 1994, Neurochem Res 19:331-338).

Lactoferrin improves neuron density and neuron survival and is able torepair and/or reverse neuronal cell damage.

A morphological examination was conducted following MR acquisition.Contiguous sections at the level of the striatum, dorsal and lateralhippocampus were collected to assess cortical and hippocampalarchitecture and white matter injury. Specific cells types were labeledusing immunohistochemistry, in order to determine specific cellularresponses. Specific labeling of neurons (NeuN), astrocytes (GFAP) andradial glia (Nestin), in conjunction with markers of white mattermyelination (MBP), was performed. The brief methodology was thefollowings:

At P7 and P21, respectively, pups from each group were deeplyanesthetized using ketalar (50 mg/ml; 0.2-0.5 ml, i.p.). Animals wereperfused intracardially with 0.9% NaCl, then 4% paraformaldehyde. Brainswere removed, weighed and postfixed in 4% paraformaldehyde overnight,then 30% sucrose for 24 h minimum, and stored at −80° C. untilsectioned. Coronal sections (10 μm) at the level of the dorsalhippocampus were cut on a cryostat (Microm Cryo-Star HM 560M, MicromInternational, Germany). Three sections at 200 μm intervals werecollected from each animal.

Immunohistochemitry: Brain tissue was processed for immunoreactivity toMBP (1:400 brand city country) using the avidin-biotin peroxidasecomplex (ABC; Vector Laboratories, Burlingame, Calif., USA). Sectionswere blocked in 4% bovine serum albumin (BSA brand city country), thenincubated with the primary antibody for 24 h at 4° C., after which theywere incubated with the secondary antibody (1:200 brand city country),then with the avidin-biotin complex (1:200, Vector Laboratories,Burlingame, Calif., USA). Sections were reacted with the chromagen,3,3-diaminobenzidine (DAB brand city country) in 0.01% hydrogenperoxide, then coverslipped.

The same protocol was used for fluorescence immunohistochemistry fornestin (1:500 brand city country), GFAP (1:400 brand city country), andNeuN (1:200 brand city country), except that sections were not incubatedin the avidin-biotin complex and DAB.

Each experimental group and their respective controls were stainedsimultaneously. When the primary antibody treatment was omitted,staining failed to occur.

Quantitative analyses were performed using MetaMorph® Imaging System(Meta Imaging Software, Molecular Devices Corporation, Pennsylvania,U.S.A). Values for each animal were pooled and a mean of means±SEM wascalculated for each group. Measurements were made on coded slidesblinded to the observer with the codes not being disclosed until theconclusion of analyses.

The results were the following and are shown in FIG. 7

The histological analysis revealed that LF supplementation Dex pup (n=5)has significant increased the Nuclei morphology and neuron density inthe CA2-CA3 field of the hippocampus compared to the Dex control pup atP7. A decrease in neuronal density in the cortex at P7 suggests neuronalloss. The neuronal density is similar to the normal control vehiclegroup (FIG. 7). Lactoferrin given in this particular developmental timeframe will influence neuronal density in the hippocampus and area ofgreat vulnerability for undernutrition and stress related brainabnormalities. This implies that LF administration increases neuronsurvival and neuron protection, for example in a young IUGR rat.

Lactoferrin supplementation increases gene expression of Brain derivedneutrophic factor (BDNF).

FIG. 8 shows that dietary lactoferrin supplementation significantlyincreased gene expression of brain derived neutrophic factor (BDNF) atpostnatal day 7.

BDNF is a neurotrophic factor that promotes neuronal differentiation,survival, and plasticity in the peripheral nervous system and centralnervous system (CNS). It is a key molecule involved in many neuronalaspects of developing and mature neurons. In CNS, BDNF elicits long-termpotentiation, which is related to synaptic plasticity. BDNF promotesneurogenesis. In particular, BDNF promotes the outgrowth of neurites andincreases the expression of synaptic proteins, which are required forestablishing synaptic connections or functions during development. Thusdietary lactoferrin has a role of both neurodevelopment andneuroprotection.

Gene expression is the process by which the information encoded by agene is converted into a protein. Our study is the first to analyzeeffect of lactoferrin supplementation on gemonic analysis brain BDNFlevel using a well established method.

Briefly, total RNA from hippocampus was extracted using the RNeasy MiniKit® according to the manufacturer's protocol (Qiagen, Basel,Switzerland). 2.5 micrograms total RNA were reverse-transcribed using800 units of Moloney murine leukemia virus reverse transcriptase(Invitrogen, Basel, Switzerland), in the presence of 0.3 units/microlRNAsin (Promega Corp, Madison, Wis.), 7.5 microM of random primers(oligo(dN)6), 1.2 mM dNTP and 12 microM of DTT. The expression of thecDNAs for rat BDNF were determined by quantitative real-time PCR usingan ABI step one plus Detection System (Applera Europe, Rotkreuz,Switzerland) and were normalized using the housekeeping ribosomal gene36B4. PCR products were quantified using the SYBR Green Core Reagent kit(Applera Europe, Rotkreuz, Switzerland) and results are expressed inarbitrary units (A.U) relative to control group values. Primers weredesigned using the Primer Express software (Applera Europe, Rotkreuz,Switzerland).

Animal Behavior Data:

Bovine milk lactoferrin (bLF) was supplemented to the rat motherthroughout gestation and lactation (total 6 weeks) at a dose level of 1g/kg/day to determine the benefits of bLF on animals behavior in 4different groups: (1) control-vehicle (CE); (2) control-DEX (CD); (3)lactoferrin-vehicle (LE) and; (4) lactoferrin-Dex (LD) at the age 4.5months using an Intellicage.

The free adaptation test (relevant to open field test) was to place ratsinto the Intellicage, a new environment for 3 days to monitor the rat'smovement and interaction with the environment (number of visitsdifferent of the corners). The exploratory behavior/curiosity weremonitored to analyze how the rats adapted to the new environment.

The results showed that control DEX rats had a decreased exploratoryactivity/curiosity with the Intellicage compared to the control-vehicle(normal control), bLF-vehicle and bLF-DEX rats throughout the 3 daystrail. The differences between control-Dex and both the control-vehicleand bLf-Dex are significant at day 3 of the free adaptation trail(P<0.05). These results suggested that prenatal LF supplementationimproved the anxiety-like behavior, including more exploratory activity,curiosity and interaction to the new environment for the healthy andearly life brain injured adult animals at age 4.5 months old. Theobtained data suggest a pronounced neuron protective effect of LF and asomewhat smaller effect on neurodevelopment (FIG. 1).

The invention claimed is:
 1. A method for use in the treatment of adelayed brain development and/or a delayed development of the nervoussystem comprising administering a composition comprising lactoferrin ina concentration of 2 g-25 g/100 kcal of the composition to an individualin need of same.
 2. A method for use in the repair of a delayed braindevelopment and/or a delayed development of the nervous systemcomprising administering a composition comprising lactoferrin in aconcentration of 2 g-25 g/100 kcal of the composition to an individualin need of same.
 3. A method for use in the treatment of a delayedvision development, a delayed neural migration, and/or a delayedcognitive development comprising administering a composition comprisinglactoferrin in a concentration of 2 g-25 g/100 kcal of the compositionto an individual in need of same.
 4. A method for use in the treatmentof an impaired learning ability, an impaired mental performance, animpaired memory or a reduced attention span comprising administering acomposition comprising lactoferrin in a concentration of 2 g-25 g/100kcal of the composition to an individual in need of same.
 5. A method inaccordance with claim 1, wherein the composition is to be administeredto an individual selected from the group consisting of mothers duringpregnancy, mothers during lactation, premature or term born babies,infants, toddlers, children and teenagers.
 6. A method in accordancewith claim 2, wherein the composition is to be administered to anindividual selected from the group consisting of mothers duringpregnancy, mothers during lactation, premature or term born babies,infants, toddlers, children and teenagers.
 7. A method in accordancewith claim 3, wherein the composition is to be administered to anindividual selected from the group consisting of mothers duringpregnancy, mothers during lactation, premature or term born babies,infants, toddlers, children and teenagers.
 8. A method in accordancewith claim 4, wherein the composition is to be administered to anindividual selected from the group consisting of mothers duringpregnancy, mothers during lactation, premature or term born babies,infants, toddlers, children and teenagers.
 9. Method of claim 1, whereinthe composition is administered to provide at least 0.2 g lactoferrinper kg of body weight of the individual per day.
 10. Method of claim 1,wherein the composition is administered to provide at least 1 glactoferrin per kg of body weight of the individual per day.
 11. Methodof claim 1, wherein the individual to whom the composition isadministered has a delayed brain development and/or a delayeddevelopment of the nervous system.
 12. Method of claim 3, wherein theindividual to whom the composition is administered has a delayed visiondevelopment, a delayed neural migration, and/or a delayed cognitivedevelopment.
 13. Method of claim 3, wherein the individual to whom thecomposition is administered has an impaired learning ability, animpaired mental performance, an impaired memory or a reduced attentionspan.